A HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC ASSAY FOR CIPROFLOXACIN HYDROCHLORIDE OPHTHALMIC SOLUTION

TF01

Edith Cristina Laignier Cazedey(1), Débora Pessôa Perez(2), Hérida Regina Nunes Salgado(1,2)

(1)Programa de Pós-graduação em Ciências Farmacêuticas; (2)Departamento de Fármacos e Medicamentos - Universidade Estadual Paulista – UNESP, Faculdade de Ciências Farmacêuticas, Araraquara (SP), Brasil.

INTRODUCTION Ciprofloxacin is a fluoroquinolone antibiotic with a broad spectrum of activity used in the prophylaxis and treatment of a wide range of bacterial infection. Ciprofloxacin is the most widely prescribed fluoroquinolones in the world, followed by ofloxacin. The objective of the current study was to development and subsequent validate of a simple, sensitive and precise reversed-phase HPLC method for the determination of ciprofloxacin hydrochloride in ophthalmic solution form. EXPERIMENTAL Ciprofloxacin hydrochloride reference substance (assigned purity 100%) was supplied by EMS Sigma Pharma group, Brazil. Ciprofloxacin hydrochloride ophthalmic solutions were obtained commercially. The ophthalmic solutions were claimed to contain 3.5 mg mL^-1 of drug and boric acid, sodium citrate, dissodium edetate (EDTA), benzalconium chloride and water as excipients. The mobile phase consisted of acetic acid 2.5%, methanol and acetonitrile (70:15:15, v/v/v). High purity water Milli-Q was used as solvent. The liquid chromatographic system, used in the present study, consisted of a Waters chromatograph equipped with Waters 1525 pump binary grade, Rheodyne Breeze 7725i injection valve with a 20 µL loop and UV-Vis Waters 2487 detector. The separation was performed at temperature of 21 ± 2 ºC, on a reversed phase Symmetry Waters C₁₈ column (250 mm x 4.6 mm i.d., 5 µm particle size). UV detection of the analyte was carried out at 275 nm. The flow rate of 1.5 mL min^-1 was maintained. RESULTS AND DISCUSSION Linear calibration plots for the ciprofloxacin hydrochloride assay method were obtained over the calibration range of 1.0–6.0 µg mL^-1.The correlation coefficient obtained was 0.9994 (n = 6), with slope and intercept were 119062 and 38613, respectively. The results exhibited an excellent correlation existed between the peak area and concentration of the analyte. The precision and robustness were assessed as repeatability, intermediate precision and varying the conditions room temperature and analyst. The RSD% values of the measurements was 3.39 and 3.77% for precision and intermediate precision, respectively, and 2.02% for robustness, confirming good precision and robustness of the proposed method. The accuracy of the method was established by recovery experiments. This study was employed by addition of known amounts of ciprofloxacin hydrochloride to the samples. The recovery experiments for ciprofloxacin hydrochloride show medium recovery of 100.11%. Specificity was described by ability of the method to measure the ciprofloxacin hydrochloride response in the presence of its excipients in the formulations. CONCLUSION Analytical method for ciprofloxacin hydrochloride in ophthalmic solution using high-performance liquid chromatography with ultraviolet detection was developed and validated. The method is highly sensitive, simple, precise, accurate and selective in pharmaceutical preparations. The results indicated that the proposed method may be recommended for determining ciprofloxacin hydrochloride in ophthalmic solutions. (This work was supported by the CAPES, CNPq and Fapesp program).